The nicotinic alpha subunit MARA1 is necessary for cholinergic evoked calcium transients in Manduca neurons.

The functional contribution of cloned subunits to insect nicotinic acetylcholine (ACh) receptors has been difficult to determine using heterologous expression. Instead, in this study we explore the subunit composition of naturally expressed functional receptors in an insect using RNA interference. The nicotinic alpha subunit, Manduca ACh Receptor Alpha 1 (MARA1) can be detected in neuronal cultures isolated from the ventral nerve cord of fifth instar larvae of Manduca sexta by in situ hybridization. It's presence correlates with large ACh induced, nicotinic Ca2+ responses. The expression of MARA1 is downregulated by treatment with dsRNA which significantly reduced both the number of responding cells and the amplitude of remaining Ca2+ responses. These results suggest that MARA1 is part of a nicotinic receptor functionally coupled to Ca2+ entry.